Altered migration potential and expression of chemokine receptors in BM DCs from ICSBP^-/- mice. (A-B) Seven-day cultured BM DCs were tested for in vitro chemotactic response using Transwell chambers. DCs were loaded on the upper chamber, while the indicated chemokines were added to the bottom wells. After a 3-hour incubation, cells that had migrated to the lower compartment were collected and counted. Data in panel A represent the mean number of migrated cells of triplicate cultures ± SE. (B) Migrated cells from triplicate cultures were pooled, double-stained with anti-CD11c and anti–I-A antibodies, and analyzed by FACS. (C) Total RNA was extracted from cell suspensions at various times of BM culture and subjected to RT-PCR, using specific primers. Day 0 represents the time of BM extraction. Transcript levels for the indicated chemokine receptors are shown in the top panel. Bottom panels show the densitometric analysis of RT-PCR bands normalized to β-actin levels. Results are representative of 3 separate experiments.